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Host-Cell Reactivation
==Introduction==
The term host cell reactivation HCR was first used to describe the survival of UV-irradiated bacteriophages, that were trasfected to UV-pretreated cells.〔Johnson, J., & Latimer, J. (2005). Analysis of DNA repair Using Transfection-Based Host Cell Reactivation. In P. Keohavong & S. Grant (Eds.), Molecular Toxicology Protocols (Vol. 291, pp. 321–335). Humana Press. Retrieved from http://dx.doi.org/10.1385/1-59259-840-4%3A321〕 This phenomenon was first thought to be the result of homologous recombination between both bacteria and phage, but later recognized as enzymatic repair.〔. Rupert, C. and Harm, W. (1966) Reactivation after photobiological damage. Adv. Radiat. Biol. 2, 1–81.〕〔Smith, K. C., & Martignoni, K. D. (1976). PROTECTION OF ESCHERICHIA COLI CELLS AGAINST THE LETHAL EFFECTS OF ULTRAVIOLET AND X IRRADIATION BY PRIOR X IRRADIATION: A GENETIC AND PHYSIOLOGICAL STUDY. Photochemistry and Photobiology, 24(6), 515–523. doi:10.1111/j.1751-1097.1976.tb06868.x〕〔Jones, D. T., Robb, F. T., & Woods, D. R. (1980). Effect of oxygen on Bacteroides fragilis survival after far-ultraviolet irradiation. Journal of Bacteriology, 144(3), 1179–1181.〕 Modifications of the assay were later developed, using transient expression plasmid DNA vectors on immortalized fibroblasts,〔Protić-Sabljić, M., & Kraemer, K. H. (1985). One pyrimidine dimer inactivates expression of a transfected gene in xeroderma pigmentosum cells. Proceedings of the National Academy of Sciences of the United States of America, 82(19), 6622–6626.〕 and lately on human lymphocytes.〔Athas, W. F., Hedayati, M. A., Matanoski, G. M., Farmer, E. R., & Grossman, L. (1991). Development and field-test validation of an assay for DNA repair in circulating human lymphocytes. Cancer Research, 51(21), 5786–5793.〕
The HCR assay known also as plasmid reactivation assay, indirectly monitors cellular transcriptional repair system, that is activated by the transcriptional-inhibited damage inflected by UV-Radiation into the plasmid. Given that UV-induced DNA damage is used as mutagen, the cell uses nucleotide excision repair NER pathway, that is activated by distortion of de DNA helix 〔Johnson, J., & Latimer, J. (2005). Analysis of DNA Repair Using Transfection-Based Host Cell Reactivation. In P. Keohavong & S. Grant (Eds.), Molecular Toxicology Protocols (Vol. 291, pp. 321–335). Humana Press. Retrieved from http://dx.doi.org/10.1385/1-59259-840-4%3A321〕
The Host-Cell Reactivation Assay or HCR is a technique used to measure the DNA repair capacity of cell of a particular DNA alteration. In the HCR assay the ability of an intact cell to repair exogenous DNA is measured〔McCready, S. (2014). An Immunoassay for Measuring Repair of DNA. In P. Keohavong & S. G. Grant (Eds.), Molecular Toxicology Protocols (Vol. 1105, pp. 551–564). Humana Press. Retrieved from http://dx.doi.org/10.1007/978-1-62703-739-6_38〕 The host cell is transfected with a damaged plasmid containing a reporter gene, usually luciferase, which has been deactivated due to the damage. The ability of the cell to repair the damage in the plasmid, after it has been introduced to the cell, allows the reporter gene to be reactivated. Earlier versions of this assay were based on the chloramphenicol acetyltransferase ((CAT) ) gene,〔Protić-Sabljić, M., & Kraemer, K. H. (1985). One pyrimidine dimer inactivates expression of a transfected gene in xeroderma pigmentosum cells. Proceedings of the National Academy of Sciences of the United States of America, 82(19), 6622–6626.〕 but the version of the assay using luciferase as reporter gene is as much as 100-fold more sensitive.〔Johnson, J., & Latimer, J. (2005). Analysis of DNA Repair Using Transfection-Based Host Cell Reactivation. In P. Keohavong & S. Grant (Eds.), Molecular Toxicology Protocols (Vol. 291, pp. 321–335). Humana Press. Retrieved from http://dx.doi.org/10.1385/1-59259-840-4%3A321〕
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